101 Pak Vet J, 2010, 30(2): 100-104.culture isolation of organism. Recently, new tests have separated in another glass tube without EDTA to collectbeen developed like gamma interferon ELISA but these serum. The haematological studies were carried out astests are quite expensive to be conducted routinely in described by Benjamin (1978). Serum total proteins wereresource poor countries. These tests are reported to be determined by the Biuret method, as described by Oserused in conjunction with tuberculin test for more accurate (1976). The serum albumin was determined following thediagnosis. In the absence of these tests for poor countries, method of Varley et al. (1980). The globulin was estimatedthe use of haemato-biochemical parameters could be of by subtracting serum albumin from serum total proteins.value in enhancing the diagnostic accuracy. To have the The results obtained were subjected to statisticalidea of its usefulness in different animal species, the analysis by using General Linear Model procedure andpresent work was conducted involving a larger sample means were compared by Dunett’s test on personalsize in buffaloes, cattle, sheep and goats along with computer using SAS statistical software (SAS, 2004).comparative tuberculin test. Correlation statistics was also computed. MATERIALS AND METHODS RESULTS A total of 2057 buffaloes, 2273 cattle, 4983 sheep In buffaloes, the results on haematological parametersand 1987 goats were tested using the single comparative showed significantly lower red blood cell countintradermal tuberculin test at the neck region, using (P=0.0026), white blood cell count (P<0.0001),bovine (50,000 IU/ml) and avian (25,000 IU/ml) PPDs haemoglobin (P<0.0001) concentration, neutrophils andproduced at the authorized laboratory at the Istituto eosinophils (P=0.01), while monocyte percentage wasZooprofilattico Umbria e Marche, Italy. These animals higher in positive reactor than negative reactor animalswere from Government Livestock Experiment Stations of (Table 1). Serum total proteins, albumin and globulinsPunjab and villages of two districts of Punjab. Buffalo and were significantly (P<0.0001) higher in positive reactorcattle studied in two cities were from the same locality as animals (Table 1).for sheep and goats, while a total of 15 Government In cattle, haematological parameters revealedlivestock farms were included in the study. The negative significantly lower values of red blood cell countanimals included were from small holdings at villages and (P=0.005), packed cell volume (P<0.0001) and neutrophilsmall number was taken from a farm where animals were percentage (P=0.0004), while lymphocyte percentagenewly purchased and were tested negative. The protocols (P=0.0003) was significantly higher in positive reactorfor the production of PPDs, the execution of the test and cattle (Table 2). Among serum proteins, only serumits interpretation were carried out according to the criteria albumin was significantly lower (P=0.02) in positivedescribed in the OIE Manual of Standards for Diagnostic reactor cattle (Table 2).Tests and Vaccines (OIE, 2004). In sheep, only packed cell volume (PCV) differed Blood was collected from all the positive reactors significantly between positive and negative reactorincluding, 137 buffaloes, 134 cattle, 36 sheep and 33 animals, while all other parameters of haematology andgoats. Almost same numbers of negative animals of about serum proteins did not differ statistically (Table 3). PCVsame status (age, parity etc.) were also included and blood percentage was lower (P=0.02) in positive reactor sheepsamples were collected from these animals (140 buffaloes, (Table 3). Similarly, haematological parameters and130 cattle, 36 sheep and 33 goats). About 15 ml of whole serum proteins in goats did not differ statistically betweenblood was collected from jugular vein of each animal. Out positive and negative reactor animals, except basophilof this, 5 ml was separated in a clean glass tube having percentage which was higher (P=0.002) in positive reactorsufficient quantity of EDTA and 10 ml of blood was animals (Table 4).Table 1: Comparison of mean haematological parameters and serum proteins in tuberculin positive and negative reactor buffaloes Parameters studied Negative animals (n = 140) Positive reactors (n = 137) P value RBC (106/µl) 5.77 ± 1.34 5.09 ± 1.81 0.002 WBC (103/µl) 5.88 ± 2.89 2.59 ± 3.44 <0.0001 Hb (g/dl) 13.86 ± 6.34 11.05 ± 2.07 <0.0001 PCV (%) 31.76 ± 4.59 32.91 ± 7.14 ESR (mm/hr) 79.54 ± 43.52 77.24 ± 44.17 Differential leukocyte count Neutrophils (%) 40.82 ± 7.30 38.05 ± 8.62 0.01 Lymphocytes (%) 48.80 ± 7.30 50.62 ± 9.26 Monocytes (%) 3.92 ± 2.69 5.99 ± 3.35 <0.0001 Eosinophils (%) 6.40 ± 2.61 5.26 ± 3.99 0.01 Basophils (%) 0.27 ± 0.58 0.41 ± 1.11 Serum proteins Serum total proteins (g/dl) 7.55 ± 1.61 10.18 ± 3.22 <0.0001 Serum albumin (g/dl) 3.77 ± 0.82 4.73 ± 1.60 <0.0001 Serum globulins (g/dl) 3.55 ± 0.88 5.07 ± 2.61 <0.0001
103 Pak Vet J, 2010, 30(2): 100-104.Correlation with severity, acuteness or chronicity of the disease. The results of correlation statistics were different in Previously, a drop in red blood cell count and haemoglo-positive and negative reacting animals of each species. A bin concentration (Samad and Rahman, 1986; Amin,significant positive correlation of haemoglobin with red 1989; Rao et al., 1992; Kumar et al., 1994), PCV (Kumarblood cell count and PCV in positive reacting buffaloes et al., 1994) and neutrophils (Samad and Rahman, 1986)(r=0.348, P<0.0001; r=0.438, P<0.0001), goats (r=0.805, has been reported in positively reacting cattle. TheP<0.001; r=0.424, P =0.02) and sheep (r=0.731, increase in monocytes (in buffaloes) and lymphocytes (inP<0.0001; r=0.353, P =0.03) was observed, while this cattle) correlated to some extent with earlier finding incorrelation was non-significant in negative animals of bison, where these were slightly higher than controlthese species. The correlation statistics for differential animals (Miller et al., 1989). A significant decrease inleukocyte percentages did not differ between positive and leukocytes in buffaloes, while non-significant decrease innegative reactor animals of these species. cattle, was another supportive finding to tuberculin results The correlation statistics of serum proteins revealed that has been previously reported and related withsimilar findings in cattle and goats with albumin showing peracute to acute form of the disease (Lepper et al., 1977).significant positive correlation with globulins in Similarly, increase in lymphocyte percentage and alsonegatively reacting cattle (r=0.413, p=0.01) and goats total leukocyte count (contrary to our results) has been(r=0.745, p<0.0001), while this correlation was non- reported (Samad and Rahman, 1986). Kumar et al. (1994)significant in positive reactor animals of these species. also reported that depending on the stage and progress ofHowever, in buffaloes, albumin showed a significant the disease, leukocytosis, leucopoenia, lymphocytosis,positive correlation with globulins (r=0.255, p=0.03), lymphopenia, neutrophilia and monocytosis can bewhile this correlation was non-significant in negative observed. Our results also confirm the earlier findingsreactor buffaloes. (Kumar et al., 1994) about no utility of ESR in animals in the diagnosis of tuberculosis or supportive diagnosis of DISCUSSION tuberculin testing and thus our results disagree with the increase in ESR in positive animals reported earlier Haematological and serum protein variations in 18 (Samad and Rahman, 1986; Amin, 1989).tuberculin positive buffaloes have previously been The results on these parameters in both sheep andreported (Javed et al., 2006), while the data about goats were almost similar with non-significant differenceepidemiological factors in sheep and goats have been between positive and negative reactor animals, withpresented in another paper (Javed et al., 2010). This study exception of PCV which was significantly lower in sheep,reports data of 137, 134, 36 and 33, positive reacting while non-significantly in goats. This suggests that thesebuffaloes, cattle, sheep and goats, respectively to avian species behave differently than large ruminants. However,and bovine PPD. Therefore, this data gives a better picture a significantly lower PCV in sheep was also observed inabout haematological and serum proteins variations positive cattle, thus supporting the results for thisamong tuberculin positive and negative reacting animals parameter to be of some utility in sheep and cattle inunder tropical conditions. conjunction with results of tuberculin test. The results of haematological and serum proteins The correlation statistics appears to have some moreparameters in buffaloes and cattle show species meaningful results in multi animal species. A significantdifferences. However, some of the results were similar in positive correlation existed of Hb with RBC and PCV inboth species like the decrease in red blood cell count and three species of animals (buffaloes, goats and sheep),neutrophil percentage. Positive reactor buffaloes showed while a significant negative correlation of albumin withsignificant decrease in RBCs, WBCs, Hb concentration globulins was observed in two species (cattle and goats).and eosinophils, while monocytes percentage along with The latter correlation in buffaloes was also significant butserum total proteins, albumin and globulins showed positive. This suggests usefulness of the correlationsignificant increase. However, in cattle, lymphocyte statistics along with haematology and serum proteins inpercentage was higher, while PCV and serum albumin strengthening the results of PPD testing in these species.were lower in positive animals. This suggests a species This kind of data would be very helpful in the diagnosis ofvariation. However, the results indicate that RBC count tuberculosis along with tuberculin testing in resource poorand neutrophil percentage can be taken as subjective countries where advanced diagnostic facilities are costlyindicator along with results of PPD testing for and mostly unavailable and animal test and slaughterconfirmation of tuberculosis in these species. The policy along with strong abattoir vigilance is missing tomonocyte percentage in buffaloes and lymphocyte confirm the tuberculin results. Further, the results alsopercentage in cattle can be considered supportive suggest that buffaloes behave differently than cattle andindicators of disease in these species. There was however, these parameters (haematology and serum proteins) area clear variation in serum proteins pattern in both the probably of less significance in small ruminants.species with increase in buffalo and decrease in cattle.The results on studied parameters can however, vary with Conclusionsthe stage of the disease in both the animal species (Lepper It can be concluded from the present study thatet al., 1977). We were unable to find any clinical sign tuberculosis causes decrease in erythrocyte parametersindicative of tuberculosis in these animals nor we had any including RBC count, PCV and Hb concentration, while itsuch history, therefore, experimental studies are needed to causes an increase in monocytes, eosinophils andbetter correlate these results in both buffaloes and cattle lymphocytes. Further, the results of haemato- logy need to
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